Modified cell-electrospinning for 3D myogenesis of C2C12s in aligned fibrin microfiber bundles
Modified cell-electrospinning for 3D myogenesis of C2C12s in aligned fibrin microfiber bundles
复制标题
DOI：
10.1016/j.bbrc.2019.06.082
复制
发表时间：
2019-08-20
影响因子：
3.1
通讯作者：
Grayson, Warren L.
中科院分区：
文献类型：
--
作者：
Guo, Yanheng;Gilbert-Honick, Jordana;Grayson, Warren L.
电纺方法可以产生具有排列线索的支架，以指导肌源性前体细胞发展成为3D骨骼肌移植物。然而，种植在这些支架上的细胞附着在支架的外部，导致支架内部的无细胞区域。为了克服这一局限性，我们改进了一种水溶液-电纺丝方法来包裹C2C12，并将其电纺成纤维蛋白/聚氧化乙烯(PEO)微纤维束。我们证明，负载C2C12作为细胞聚集体(直径80-90微米)并改变其他几个电纺参数可以显著提高暴露在4.5千伏电场下的细胞存活率。C2C12种子纤维蛋白/PEO微纤维束培养长达7天。未诱导和成肌诱导的C2C12细胞增殖、延长并成为多核细胞。肌管长度(122.4±10.9微米比59.9±8.3微米)和肌管直径(16.76±2.06微米比12.49±0.93微米)增加。这项研究中提供的数据首次表明，在水溶液静电纺丝过程中，细胞可以被加载到排列的纤维蛋白水凝胶3D结构中，同时保持其形成新生组织的潜力。(C)2019 Elsevier Inc.保留所有权利。
Electrospinning methods can generate scaffolds with alignment cues to guide the development of myogenic precursors into 3D skeletal muscle grafts. However, cells seeded onto these scaffolds adhere to the exterior resulting in regions of acellularity within the scaffold interior. To overcome this limitation, we modified an aqueous solution-electrospinning method to encapsulate C2C12s and electrospin them into fibrin/polyethylene oxide (PEO) microfiber bundles. We demonstrated that loading C2C12s as cellular aggregates (80-90 mu m in diameter) and modifying several other electrospinning parameters dramatically increased cell viability following exposure to the 4.5 kV electric field. C2C12-seeded fibrin/PEO microfiber bundles were cultured for up to seven days. Uninduced and myogenically induced C2C12s proliferated, elongated and became multinucleated. Myogenic induction increased the number of myotube-associated nuclei (36.4 +/- 12% vs. 6.2 +/- 1.9%), myotube length (122.4 +/- 10.9 mu m vs. 59.9 +/- 8.3 mu m), and myotube diameter (16.76 +/- 2.06 mu m vs. 12.49 +/- 0.93 mu m). The data presented in this study demonstrates for the first time that cells can be loaded inside the aligned fibrin hydrogel 3D construct during aqueous solution electrospinning while retaining their potential for de novo tissue formation. (C) 2019 Elsevier Inc. All rights reserved.